Sains Malaysiana 50(5)(2021): 1393-1405

http://doi.org/10.17576/jsm-2021-5005-18

Anti-Tumor Activity of Metformin in Human Epidermal Growth Factor Receptor 2 Positive Breast Cancer Cells

(Aktiviti Anti-Tumor Metformin pada Reseptor Faktor Pertumbuhan Epidermis Manusia 2 Positif Sel Kanser Payudara)

FATHUL HUDA^1,2*, SARI EKAWATI³, ANINDY PUTRI ADDINA³, AHMAD FARIED^2,4, AFIAT BERBUDI^1,2, TAOFIK RUSDIANA⁵, TENNY PUTRI⁶, NURUL QOMARILLA⁶, LUKMAN HILFI⁷, IWAN SETIAWAN¹ & MUHAMMAD HASAN BASHARI^1,2

¹Department of Biomedical Science, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia

²Oncology and Stem Cell Working Group, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia

³Bachelor of Medicine Program, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia

⁴Department of Neurosurgery, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia

⁵Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Bandung 40161, Indonesia

⁶Cell Culture Laboratory, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia

⁷Department of Public Health, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia

Received: 18 December 2019/Accepted: 29 September 2020

ABSTRACT

Breast Cancer (BC) is the leading cause of cancer death in women. One BC subtype is very aggressive with amplification of human epidermal growth factor receptor 2 (HER2) protein. Although specific HER2+ targeting agents are available, most of HER2+ BC patients develop resistant to these agents. Recent studies show that metformin, is able to become anti-tumor in various cancer cells. This research aims to evaluate anti-tumor activities of metformin to HER2+ BC cells in both sensitive and resistant to trastuzumab. A series of assays were performed to evaluate metformin anti-tumor activities in HCC-1954 and SKBR-3 HER2+ BC cells. MTT assay was performed to evaluate cell death, and inhibitory concentration (IC₅₀), while scratch assay was performed to assess inhibition of cell migration and clonogenic assay to assess cell proliferation. p<0.05 was considered to be significant. Metformin could suppress the number of HER2+ BC cells. Viability assay showed suppression of viable cells after metformin incubation of 60 and 600 µM compared to control, 30 and 90%, respectively. Surprisingly, IC₅₀ of metformin was smaller in HER2+ BC HCC-1954 cells that resistant to trastuzumab compare to the sensitive one (SKBR-3). Both were below 1 µM, with R² more than 0.95. Additionally, clonogenic assay showed less colony number and colony area with at least p < 0.05 in colony number and p < 0.01 in the area. In addition, metformin inhibited cell migration of HER2+ BC cells. Metformin shows a potency as anti-tumor by inducing cell death, inhibiting cell proliferation and cell migration of HER2+ BC cells.

Keywords: Breast Cancer; HER2+; metformin; Trastuzumab resistant

ABSTRAK

Kanser Payudara (BC) adalah penyebab utama kematian akibat barah pada wanita. Satu subjenis BC sangat agresif dengan penguatan protein reseptor 2 faktor pertumbuhan epidermis manusia (HER2). Walaupun agen sasaran HER2 + khusus ada, kebanyakan pesakit HER2 + BC mengalami ketahanan terhadap agen ini. Kajian terbaru menunjukkan bahawa metformin mampu menjadi anti-tumor pada pelbagai sel barah. Penyelidikan ini bertujuan untuk menilai aktiviti anti-tumor metformin kepada sel HER2 + BC dengan sensitif dan tahan terhadap trastuzumab. Satu siri ujian dilakukan untuk menilai aktiviti anti-tumor metformin pada sel HCC-1954 dan SKBR-3 HER2 + BC. Ujian MTT dilakukan untuk menilai kematian sel dan kepekatan penghambatan (IC₅₀), sementara ujian awal dilakukan untuk menilai penghambatan penghijrahan sel dan pengujian klonogenik untuk menilai percambahan sel. p <0.05 dianggap signifikan. Metformin dapat menekan bilangan sel HER2 + BC. Ujian daya maju menunjukkan penekanan sel yang berdaya maju setelah inkubasi metformin 60 dan 600 µM berbanding kawalan, masing-masing 30 dan 90%. Anehnya, IC50 metformin lebih kecil pada sel HER2 + BC HCC-1954 yang tahan terhadap trastuzumab dibandingkan dengan sel sensitif (SKBR-3). Kedua-duanya berada di bawah 1 µM, dengan R2 lebih daripada 0.95. Selain itu, ujian klonogenik menunjukkan bilangan koloni dan kawasan koloni yang kurang dengan sekurang-kurangnya p <0.05 pada bilangan koloni dan p < 0.01 di kawasan koloni. Sebagai tambahan, metformin menghalang penghijrahan sel HER2 + BC sel. Metformin menunjukkan potensi sebagai anti-tumor dengan mendorong kematian sel, menghambat percambahan sel dan penghijrahan sel HER2+ sel BC.

Kata kunci: HER2 +; kanser payudara (BC); metformin, rintangan Trastuzumab

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*Corresponding author; email: fathul@unpad.ac.id